Chemicon公司的CpGenomeDNAModificationKit的protocol修正

我用了快4个月这个试剂盒了,下面是一些总结出来的经验,我现在诱变每次都很成功了,呵呵:)1. 我溶NaOH的水PH值大概在6.5左右,所以我每次调Reagent I的时候3MNaO0H基本都要加150uL左右。2. 解链时用50度水浴15分钟。3. 加入Reagent I后,50度水浴16小时。4. 加入Reagent II和III后,室温孵育10分钟5. 第一次离心用9000 rpm 5分钟。6. 70%乙醇清洗3次,每次9000rpm 1 分钟,最后一次13000rpm2min。7. 加入20mM NaOH/90%EtOH后,反映5分钟。8. 不要去掉20mM NaOH/90%EtOH,直接加入90%乙醇后离心,9000rpm 1min9. 90%乙醇清洗2次,每次9000 rpm 1 min,最后一次13000rpm 3min10. PH 7.5 TE buffer 65度 溶解 15 minPS: 如果发现PCR结果中M......阅读全文

Western-Blot-Protocol

一、提取抗原蛋白将提取RNA途中留存的样品,加入150μl 100%酒精充分混匀,静置5min(RT), 2000×g , 4℃离心5min, 吸取上清至新管中, 加入750μl异丙醇, 混匀, 静置10min(RT), 12000×g, 4℃离心10min, 弃上清, 加入1ml 0.3mol/L

Protocol-for-dsRNA-Synthesis

实验概要        We routinely produce dsRNA by in vitro transcription of a PCR generated DNA template containing the T7 promoter sequence on both ends

Microarray-Hybridization-Protocol

Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an

Phycoerythrin-conjugation-protocol

Phycoerythrin conjugation protocolDavid's method modified from references (2) and (3). I used this method to conjugate a mouse IgG2a monoclonal an

Urea-Lysis-Protocol

Urea lysis buffer            9M Urea, 2.5mM EDTA, 2.5mM EGTA, 1% DTE, 4% CHAPS            make 10ml and aliquot 10x1ml, freeze at -70°C Lysate prepara

Tissue-Harvest-Protocol

TISSUES TO BE PROCURED(minimally and preferably within 5-8 hours after death):1. Brain2. Liver3. Muscle4. Skin5: Others as the specific case dictatesP

RNA-Isolation-Protocol

RNA Isolation Protocol(Revised 5-15-2003)Stabilize RNAStart with 15 ml E. coli Culture containing 7.5* 109 cells (OD600= 0.2 Dilute cells or scale up)

Protocol-for-Cell-Fusion

Healthy Sp2/0 cells should be rapidly growing by this time. Sp 2/0 cells should be started about two weeks before the cell fusion. Every two days, the

Yale-Immunofluorescence-Protocol

实验概要We provide a protocol for fixation, immunostaining, and imaging in 384-well Plates.主要试剂Reagents1. 384-well view plates (Aurora)2. HUVEC (pooled, L

Dot-blot-protocol

实验概要A  technique for detecting, analyzing, and identifying proteins, similar  to the western blot technique but differing in that protein samples are

Nucleolar-Isolation-Protocol

We recommend that you first download and read this page as a PDF file. Using that as your guide, you can then follow the protocol below and view a Qui

Protocol-for-Trichl...

实验概要The  efficiency of nucleotide incorporation in DNA/RNA polymerization  reactions (e.g. transcription, reverse transcription, and DNA  replication)

Western-Blotting-Protocol

实验概要The western blot  (sometimes called the protein immunoblot) is a widely used analytical  technique used to detect specific proteins in the given s

外显子基因为什么保守?《Nature-Genetics》出新答案了!

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流式细胞仪技术专辑

Flow Cytometry Analysis (Springer Lab, Harvard University) Flow cytometry employs instrumentation that scans single cells flowing past excitation sour

Human-DC-Enrichment-Kit

实验概要Enrich  untouched Dendritic Cells (DCs) by depleting T cells, B cells,  monocytes/ macrophages, NK cells, erythrocytes and most granulocytes  from

Dynabeads®-Streptavidin-Trial-Kit

实验概要Intended UseDynabeads®  Streptavidin are ideal for numerous applications, including  purification of proteins, nucleic acids purification, protein

Dynabeads®-Antibody-Coupling-Kit

实验概要Non-specific  binding of an antibody to proteins other than the antigen can sometimes  occur. This is usually more common with polyclonal antibodi

Dynabeads®-Antibody-Coupling-Kit

实验概要The  Dynabeads and buffers provided in this kit will enable you to  covalently immobilize antibodies (or other protein ligands such as  lections,

PCR基本实验方法(四)

Labelling PCR Products with DigoxigeninPCR products may be very conveniently labelled with digoxigenin-11-dUTP (Boehringer-Mannheim) by incorporating

PCR基本实验方法(四)

 Labelling PCR Products with DigoxigeninPCR products may be very conveniently labelled with digoxigenin-11-dUTP (Boehringer-Mannheim) by incorporating

CellTrace™-CFSE-Cell-Proliferation-Kit

实验概要The CellTrace™  CFSE Cell Proliferation Kit provides a versatile and well-retained  cell-tracing reagent in a convenient and easy-to-use form. The

Infusion-biobrick-assembly

OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are cur

美DNA检测公司不再提供健康信息

美国基因检测公司“23和我”已经暂停提供个人健康信息   近日,“23和我”公司宣布,将不再为购买其基因试剂盒的顾客提供健康信息。该公司发布的声明表示,这一变化是“遵照美国食品药品监督管理局(FDA)的新指令,在我们接受监管审查的过程中,停止新的消费者使用”。“23和我”是一家基因测试公司,总部位

DNA测序最大赢家:Illumina公司

  对包括生物科技投资者在内的大多数人而言,人类基因工程是一大失败,未能带来新药物、医学新突破和其他的实际好处。因此,在我开始发表我称为“基因周”的系列博文之时,让我们回顾一下所有医学类股票中表现最佳者,其中不仅涉及生物科技,还涉及医疗设备和药物。生物科技领域里的最大赢家们公司5年总回报5

流式细胞仪技术专辑

 最方便的实验干货查询工具微信扫码进入「丁香实验」小程序编辑: 呜咽分享到:      Flow Cytometry Analysis (Springer Lab, Harvard University)Flow cytometry employs instrumentation that scan

SYBR-Green-Quantitative-PCR-Protocol

SummaryQuantitative PCR is a method used to detect relative or absolute gene expression level. All qPCR involves the use of fluorescence to detect the

免疫荧光

Immunofluorescence Technique (Spector Lab)protocol for immunofluorescence on cells  Immunofluorescence Protocol (Walter Steffen)Methanol fixationForma

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电子天平的修正方法1、当水平泡处于水平圈的上方时,表明电子天平的上方位置偏高,我们要通过旋转电子天平的上部的两个转角,使泡泡下移。2、当水平泡处于水平圈的下方时,表明电子天平的下方位置偏高,我们要通过旋转电子天平的下部的两个转角,使泡泡上移。3、当水平泡处于水平圈的左侧时,表明电子天平的左侧位置偏高

涂层测厚仪基本校准的修正

在下述情况下,改变基本校准是有必要的:____测头顶端被磨损。____特殊的用途。在测量中,如果误差明显地超出给定范围,则应对测头的特性重新进行校准称为基本校准。通过输入 6个校准值(1 个零和5 个厚度值),可重新校准测头。a) 在仪器关闭的状态下按住“↑”键再按“ON/OFF”键,即进入F 型测